Figure 2: Effects of pcDNA3.1-HBx on the expression of AFP, Src, CXCR4, Ras, pAKT(Ser473), and p-mTOR(Ser2448) in liver cells(A) L-02 and CHL liver cells were transfected with pcDNA3.1-HBx and expression of HBx was measured 48 hours later by Western blotting. (B) Expression of Src, CXCR4, Ras, pAKT(Ser473), and p-mTOR(Ser2448) was measured by Western blotting 1, 7, 14, 21, and 28 days after transfection. Results are from one representative experiment of three.

Mentions: We transfected a vector expressing the HBV protein HBx, pcDNA3.1-HBx, into human liver cell lines L-02 and CHL and measured the impact of pcDNA3.1-HBx on the expression of AFP and the oncogenes Src, CXCR4, and Ras. The pcDNA3.1-HBx-mediated induction of HBx expression in L-02 and CHL cells was evident 2 days after transfection and remained elevated between 7 and 28 days after transfection (Figure 2A). Expression of AFP was emerged after transfected with pcDNA3.1-HBx for 7 days and persisted increasing after 28 days. Expression of CXCR4 and Ras was enhanced 7 day after transfection and increased for 28 days in both L-02 and CHL cells (Figure 2B). The expressions of AFP and CXCR4 were further confirmed at the mRNA levels by quantitative RT-PCR analysis (Supplementary Figure 1B). Src expression was elevated 14 days after transfection in both L-02 and CHL cells (Figure 2B).